Vascular casted prostheses and methods of forming same for treating biological tissue

ABSTRACT

Bioremodelable tissue prostheses having a vasculature that is infused with a biomaterial composition.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Application No. 62/088,987,filed on Dec. 8, 2014.

FIELD OF THE INVENTION

The present invention relates to methods and apparatus for repairingdamaged or diseased biological tissue. More particularly, the presentinvention relates to non-antigenic, resilient, bioremodelable tissueprostheses having a vasculature that is infused with a biomaterialcomposition.

BACKGROUND OF THE INVENTION

As is well known in the art, tissue prostheses are often employed totreat or replace damaged or diseased biological tissue. However, despitethe growing sophistication of medical technology, the use of prosthesesto treat or replace damaged biological tissue remains a frequent andserious problem in health care. The problem is often associated with thematerials employed to construct the prostheses.

As is also well known in the art, the optimal prosthesis material shouldbe chemically inert, non-carcinogenic, capable of resisting mechanicalstress, capable of being fabricated in the form required, andsterilizable. Further, the material should be resistant to physicalmodification by tissue fluids, and not excite an inflammatory reaction,induce a state of allergy or hypersensitivity, or, in some cases,promote visceral adhesions. See, e.g., Jenkins, et al., Surgery, vol.94(2), pp. 392-398 (1983).

Various materials and/or structures have thus been employed to constructprostheses that satisfy the aforementioned optimal characteristics,including tantalum gauze, stainless mesh, Dacron®, Orlon®, Fortisan®,nylon, knitted polypropylene (e.g., Marlex®), microporousexpanded-polytetrafluoroethylene (e.g., Gore-Tex®), Dacron reinforcedsilicone rubber (e.g., Silastic®), polyglactin 910 (e.g., Vicryl®),polyester (e.g., Mersilene®), polyglycolic acid (e.g., Dexon®),processed sheep dermal collagen, crosslinked bovine pericardium (e.g.,Peri-Guard®), and preserved human dura (e.g., Lyodura®).

As discussed in detail below, although some of the noted prosthesismaterials satisfy some of the aforementioned optimal characteristics,there are several disadvantages and drawbacks associated with thematerials.

The major advantages of metallic reinforced prostheses, e.g., stainlesssteel and Nitinol® meshes, are that they are inert, resistant toinfection and can stimulate fibroplasia. Several major disadvantages arefragmentation, which can, and in many instances will, occur after thefirst year of administration, and the lack of malleability.

Further, many conventional prostheses; particularly, stents are oftenconstructed from various polymeric materials, such as poly(ethyleneterephthalate) (PET). Such prostheses often cause irritation andundesirable biologic responses from the surrounding tissues in a vessel.

Although conventional prostheses are designed to be implanted for anextended period of time, it is sometimes necessary to remove the deviceprematurely, for example, because of poor patency or harsh biologicalresponses. In such instances, the device generally must be removedthrough a secondary surgical procedure, which can, and in many instanceswill, result in undesirable pain and discomfort to the patient andpossibly additional trauma to the vessel tissue. In addition to the painand discomfort, the patient must be subjected to an additional timeconsuming and complicated surgical procedure with the attendant risks ofsurgery.

More recently, bioabsorbable and/or biodegradable prostheses have beendeveloped in an effort to eliminate the harsh biological responsesassociated with conventional polymeric and metal vascular prostheses.There are, however, several known disadvantages associated withbioabsorbable and biodegradable prostheses.

One major disadvantage is that the bioabsorbable and biodegradablematerials and, hence, prostheses often break down at a faster rate thanis desirable for the application. A further disadvantage is that thebioabsorbable and biodegradable materials can, and in many instanceswill, break down into large, rigid fragments that can cause obstructionsin the interior of a vessel.

A further disadvantage associated with conventional prostheses is thatexisting means for securing the prosthesis into or onto biologicaltissue within a body vessel have had limited success. Often the securingmeans comprises engaging the prosthesis to the surrounding tissue byphysical or mechanical means, such as disclosed in U.S. Pat. No.7,918,882. Another securing means comprises modifying the prosthesissurface or material to induce the production of fibrous (scar) tissue toanchor the prosthesis upon implantation within the vessel.

Various polymer based apparatus have also been developed in an attemptto construct reinforced prostheses. Illustrative are the ECM and polymerbased apparatus, i.e. grafts and endografts, disclosed in U.S. Pat. Nos.6,015,432 and 8,142,506.

U.S. Pat. No. 6,015,432 discloses a fiber-reinforced hydrogel prosthesisthat is intended to replace cartilaginous materials, wherein the fiberscomprise a polymeric material, such as polyurethane fibers.

U.S. Pat. No. 8,142,506 discloses an endovascular tube or bifurcatedprosthesis comprising a polymeric material reinforced by a threadedsuperelastic metal wire, such as a Nitinol®.

A major drawback of the noted polymer based apparatus, as well as mostknown apparatus, is that the apparatus often comprise or include apermanent structure that remains in the body, i.e. non-biodegradable. Asis well known in the art, such structures (or devices) can, and in mostinstances will, cause irritation and undesirable biologic responses inthe surrounding tissue.

Such structures (and devices) are also prone to failure, resulting insevere adverse consequences, e.g., ruptured vessels.

There is thus a need to provide improved prostheses that substantiallyreduce or eliminate (i) intimal hyperplasia after intervention in avessel, (ii) the harsh biological responses associated withconventional, and (iii) employ effective vessel securing means.

There is also a need to provide prostheses that can replace or improvebiological functions or promote the growth of new tissue in a subject.

There is also a need to provide prostheses that substantially reduce oreliminate the formation of inflammation and infection.

There is also the need to provide prostheses having mechanicalcompatibility or enhanced mechanical properties. As is well known in theart, a mismatch between the stiffness, hardness, and porosity of aprosthesis in comparison to the surrounding tissue environment can causeirritation and other complications after implantation.

It is therefore an object of the present invention to provide prosthesesthat substantially reduce or eliminate (i) intimal hyperplasia afterintervention in a vessel, (ii) the harsh biological responses associatedwith conventional polymeric and metal prostheses, (iii) employ effectivevessel securing means, and (iv) the formation of biofilm, inflammationand infection.

It is another object of the present invention to provide prostheses thatcan effectively replace or improve biological functions or promote thegrowth of new tissue in a subject.

It is another object of the present invention to provide prostheses thatinclude effective reinforcing means for temporarily positioning theprostheses proximate target tissue and retaining strength.

It is another object of the present invention to provide prostheses thatcan administer one or more pharmacological or therapeutic agents to asubject.

SUMMARY OF THE INVENTION

The present invention is directed to casted constructs for treating,reconstructing or replacing damaged or diseased biological tissue.

As discussed in detail herein, in a preferred embodiment, the castedconstructs comprise an ECM member having a biomaterial compositiondisposed within the vasculature of the ECM member.

In some embodiments of the invention, the casted constructs comprise aseamless ECM material derived from a mammalian tissue source. Accordingto the invention, the seamless ECM material can be derived from variousmammalian tissue sources, including, without limitation, smallintestine, large intestine and umbilical cord.

In some embodiments of the invention, the casted constructs comprise aseamed ECM material derived from a mammalian tissue source. According tothe invention, the ECM material can similarly be derived from variousmammalian tissue sources. In some embodiments of the invention, themammalian tissue sources include, without limitation, the smallintestine, large intestine, stomach, lung, liver, kidney, pancreas,placenta, heart, bladder, prostate, tissue surrounding growing enamel,tissue surrounding growing bone, and any fetal tissue from any mammalianorgan.

In a preferred embodiment, the mammalian ECM material referenced abovecomprises sterilized acellular ECM material.

Preferably, the mammalian tissue sources referenced above comprise anadolescent mammalian tissue source.

In some embodiments, the biomaterial composition comprises anECM-mimicking biomaterial composition, such as poly(glycerol sebacate)(PGS).

In some embodiments of the invention, the biomaterial compositioncomprises a polymeric composition further comprising a biocompatiblepolymeric material, such as poly(ε-caprolactone) (PCL).

In some embodiments of the invention, the biomaterial compositioncomprises an ECM composition comprising at least one ECM material.

In some embodiments of the invention, the ECM material and/orbiomaterial composition and, hence, casted constructs formed therewithfurther comprise at least one additional biologically active agent orcomposition, i.e. an agent that induces or modulates a physiological orbiological process, or cellular activity, e.g., induces proliferation,and/or growth and/or regeneration of tissue.

In some embodiments, the biologically active agent comprises a cell,such as, without limitation, a human embryonic stem cell, fetalcardiomyocyte, myofibroblast, and mesenchymal stem cell.

In some embodiments, the biologically active agent comprises a growthfactor, such as, without limitation, a transforming growth factor-alpha(TGF-α), transforming growth factor-beta (TGF-β), fibroblast growthfactor-2 (FGF-2), basic fibroblast growth factor (bFGF), and vascularepithelial growth factor (VEGF).

In some embodiments, the ECM material and/or biomaterial composition,and, hence, casted constructs formed therewith further comprise at leastone pharmacological agent or composition (or drug), i.e. an agent orcomposition that is capable of producing a desired biological effect invivo, e.g., stimulation or suppression of apoptosis, stimulation orsuppression of an immune response, etc.

Suitable pharmacological agents and compositions include, withoutlimitation, antibiotics, anti-viral agents, analgesics,anti-inflammatories, anti-neoplastics, anti-spasmodics, andanticoagulants and/or anti-thrombic agents.

In some embodiments of the invention, the pharmacological agentcomprises a statin, i.e. a HMG-CoA reductase inhibitor, such ascerivastatin.

In some embodiments of the invention, the casted constructs furthercomprise an outer reinforcing structure.

In some embodiments of the invention, the reinforcing structurecomprises a thin member, such as a strand, that is wound about the outersurface of the casted construct.

In some embodiments of the invention, the reinforcing structurecomprises a mesh or woven structure.

In some embodiments of the invention, the reinforcing structurecomprises an ECM-mimicking biomaterial, such as PGS.

In some embodiments, the casted constructs are incubated in a maceratingagent to degrade the ECM member.

In another embodiment of the invention, there is provided a method offorming the aforementioned casted constructs of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

Further features and advantages will become apparent from the followingand more particular description of the preferred embodiments of theinvention, as illustrated in the accompanying drawings, and in whichlike referenced characters generally refer to the same parts or elementsthroughout the views, and in which:

FIG. 1 is an illustration of a mammalian small intestine vasculature;

FIG. 2 is an illustration of a mammalian small intestine vasculatureshown in FIG. 2, being infused with a biocompatible composition, inaccordance with the invention;

FIG. 3 is a perspective partial sectional view of a casted construct,showing the vasculature thereof, in accordance with the invention;

FIG. 4 is a photograph depicting a segment of resected porcine smallintestine, showing the vasculature thereof, in accordance with theinvention;

FIG. 5 is a photograph depicting a decellularized, sterile segment ofthe resected porcine small intestine, showing the vasculature thereofinfused with a tracing composition, in accordance with the invention;

FIG. 6 is a photograph depicting the full decellularized, sterileresected porcine small intestine, showing the vasculature thereofinfused with a tracing composition via anterograde injection into themain artery, in accordance with the invention;

FIG. 7 is a photograph depicting a decellularized, sterile tubularsegment of porcine small intestine, in accordance with the invention;and

FIG. 8 is a photograph depicting a decellularized, sterile resectedrabbit small intestine with the vasculature infused with a biomaterialcomposition, in accordance with the invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

Before describing the present invention in detail, it is to beunderstood that this invention is not limited to particularlyexemplified apparatus, systems, materials, compositions, structures ormethods as such may, of course, vary. Thus, although a number ofapparatus, systems, materials, compositions, structures and methodssimilar or equivalent to those described herein can be used in thepractice of the present invention, the preferred apparatus, systems,materials, compositions, structures and methods are described herein.

It is also to be understood that the terminology used herein is for thepurpose of describing particular embodiments of the invention only andis not intended to be limiting.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one having ordinaryskill in the art to which the invention pertains.

Further, all publications, patents and patent applications cited herein,whether supra or infra, are hereby incorporated by reference in theirentirety.

As used in this specification and the appended claims, the singularforms “a, “an” and “the” include plural referents unless the contentclearly dictates otherwise. Thus, for example, reference to “an active”includes two or more such actives and the like.

Further, ranges can be expressed herein as from “about” one particularvalue, and/or to “about” another particular value. When such a range isexpressed, another embodiment includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it willbe understood that the particular value forms another embodiment. Itwill be further understood that the endpoints of each of the ranges aresignificant both in relation to the other endpoint, and independently ofthe other endpoint.

It is also understood that there are a number of values disclosedherein, and that each value is also herein disclosed as “approximately”that particular value in addition to the value itself. For example, ifthe value “10” is disclosed, then “approximately 10” is also disclosed.It is also understood that when a value is disclosed that “less than orequal to” the value, “greater than or equal to the value” and possibleranges between values are also disclosed, as appropriately understood bythe skilled artisan. For example, if the value “10” is disclosed then“less than or equal to 10”, as well as “greater than or equal to 10” isalso disclosed.

Definitions

The terms “prosthesis” and “casted construct” are used interchangeablyherein, and mean and include a structure or system that is configuredfor placement on biological tissue on or in an organ, such as a lumen orvessel. As discussed in detail herein, upon placement of a biologicalprostheses or casted construct of the invention to biological tissue;particularly, damaged or diseased tissue, the casted construct induces“modulated healing”, as defined herein.

The term “biocompatible”, as used herein, means a device or materialthat is substantially non-toxic in an in vivo environment, and is notsubstantially rejected by a recipient's physiological system, i.e.non-antigenic.

The terms “cast” and “casted” are used interchangeably herein, and meanand include a structure comprising a configuration or shape provided bya cast or mold template, e.g., the vasculature of biological tissue.

The term “buffer”, as used herein, means a composition, preferably asolution that resists changes in pH when acid or alkali is added to it.Buffers typically comprise a weak acid or alkali together with one ofits salts.

The terms “vascular” and “vasculature” are used interchangeably herein,and mean and include a structure of, relating to, affecting, orcomprising a vessel or vessels; particularly, a vessel or vessels thattransport blood.

The term “kinetics” as used herein, means and refers to the rates ofchemical and/or biochemical reactions, including the rates of multiplesimultaneous or overlapping reactions, which may be directly orindirectly related, e.g. the activation of growth factors and/orcytokines.

The term “macerated” as used herein, means and refers to the gradualdegradation of a polymeric or biological composition through chemical,enzymatic and/or mechanical means.

The terms “extracellular matrix” and “ECM” are used interchangeablyherein, and mean and include a collagen-rich substance that is found inbetween cells in mammalian tissue, and any material processed therefrom,e.g. decellularized ECM. According to the invention, the ECM materialcan be derived from various mammalian tissue sources including, withoutlimitation, the small intestine, large intestine, stomach, lung, liver,kidney, pancreas, placenta, heart, bladder, prostate, tissue surroundinggrowing enamel, tissue surrounding growing bone, and any fetal tissuefrom any mammalian organ.

The ECM material can thus comprise, without limitation, small intestinesubmucosa (SIS), urinary bladder submucosa (UBS), stomach submucosa(SS), central nervous system tissue, dermal extracellular matrix,subcutaneous extracellular matrix, gastrointestinal extracellularmatrix, i.e. large and small intestines, tissue surrounding growingbone, placental extracellular matrix, omentum extracellular matrix,epithelium of mesodermal origin, i.e. mesothelial tissue, cardiacextracellular matrix, e.g., pericardium and/or myocardium, kidneyextracellular matrix, pancreas extracellular matrix, lung extracellularmatrix, and combinations thereof. The ECM can also comprise collagenfrom mammalian sources.

The terms “urinary bladder submucosa (UBS)”, “small intestine submucosa(SIS)” and “stomach submucosa (SS)” also mean and include any UBS and/orSIS and/or SS material that includes the tunica mucosa (which includesthe transitional epithelial layer and the tunica propria), submucosallayer, one or more layers of muscularis, and adventitia (a looseconnective tissue layer) associated therewith.

The ECM can also be derived from basement membrane of mammaliantissue/organs, including, without limitation, bladder, “urinary basementmembrane (UBM)”, liver, i.e. “liver basement membrane (LBM)”, andamnion, chorion, allograft pericardium, allograft acellular dermis,amniotic membrane, Wharton's jelly, and combinations thereof.

Additional sources of mammalian basement membrane include, withoutlimitation, spleen, lymph nodes, salivary glands, prostate, pancreas andother secreting glands.

The ECM can also be derived from other sources, including, withoutlimitation, collagen from plant sources and synthesized extracellularmatrices, i.e. cell cultures.

The term “angiogenesis”, as used herein, means a physiologic processinvolving the growth of new blood vessels from pre-existing bloodvessels.

The term “neovascularization”, as used herein, means and includes theformation of functional vascular networks that can be perfused by bloodor blood components. Neovascularization includes angiogenesis, buddingangiogenesis, intussuceptive angiogenesis, sprouting angiogenesis,therapeutic angiogenesis and vasculogenesis.

The terms “ECM-mimicking biomaterial”, and “ECM-mimicking material” areused interchangeably herein, and mean and include a biocompatible andbiodegradable biomaterial that induces neovascularization andbioremodeling of tissue in vivo, i.e. when disposed proximate damagedbiological tissue. The term “ECM-mimicking” thus includes, withoutlimitation, ECM-mimicking polymeric biomaterial compositions;specifically, poly(glycerol sebacate) (PGS).

The terms “biologically active agent” and “biologically activecomposition” are used interchangeably herein, and mean and include agentthat induces or modulates a physiological or biological process, orcellular activity, e.g., induces proliferation, and/or growth and/orregeneration of tissue.

The terms “biologically active agent” and “biologically activecomposition” thus mean and include, without limitation, the followinggrowth factors: platelet derived growth factor (PDGF), epidermal growthfactor (EGF), transforming growth factor alpha (TGF-α), transforminggrowth factor beta (TGF-β), fibroblast growth factor-2 (FGF-2), basicfibroblast growth factor (bFGF), vascular epithelial growth factor(VEGF), hepatocyte growth factor (HGF), insulin-like growth factor(IGF), nerve growth factor (NGF), platelet derived growth factor (PDGF),tumor necrosis factor alpha (TNF-α), and placental growth factor (PLGF).

The terms “biologically active agent” and “biologically activecomposition” also mean and include, without limitation, embryonic stemcells, mesenchymal stem cells, hematopoietic stem cells, bone marrowstem cells, bone marrow-derived progenitor cells, myosatelliteprogenitor cells, totipotent stem cells, pluripotent stem cells,multipotent stem cells, oligopotent stem cells and unipotent stem cells.The group also comprises cardiomyocytes, myoblasts, monocytes,parenchymal cells, epithelial cells, endothelial cells, mesothelialcells, fibroblasts, osteoblasts, chondrocytes, exogenous cells,endogenous cells, macrophages, capillary endothelial cells, autologouscells, xenogenic cells, allogenic cells, and cells derived from any ofthe three germ layers including the endoderm, mesoderm and ectoderm.

The terms “biologically active agent” and “biologically activecomposition” also mean and include, without limitation, the followingbiologically active agents (referred to interchangeably herein as a“protein”, “peptide” and “polypeptide”): collagen (types I-V),proteoglycans, glycosaminoglycans (GAGs), glycoproteins, cytokines,cell-surface associated proteins, cell adhesion molecules (CAM),endothelial ligands, matrikines, cadherins, immuoglobins, fibrilcollagens, non-fibrillar collagens, basement membrane collagens,multiplexins, small-leucine rich proteoglycans, decorins, biglycans,fibromodulins, keratocans, lumicans, epiphycans, heparin sulfateproteoglycans, perlecans, agrins, testicans, syndecans, glypicans,serglycins, selectins, lecticans, aggrecans, versicans, neurocans,brevicans, cytoplasmic domain-44 (CD-44), macrophage stimulatingfactors, amyloid precursor proteins, heparins, chondroitin sulfate B(dermatan sulfate), chondroitin sulfate A, heparin sulfates, hyaluronicacids, fibronectins, tenascins, elastins, fibrillins, laminins,nidogen/enactins, fibulin I, fibulin II, integrins, transmembranemolecules, thrombospondins, ostepontins, and angiotensin convertingenzymes (ACE).

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” are used interchangeably herein, and mean and includean agent, drug, compound, composition of matter or mixture thereof,including its formulation, which provides some therapeutic, oftenbeneficial, effect. This includes any physiologically orpharmacologically active substance that produces a localized or systemiceffect or effects in animals, including warm blooded mammals, humans andprimates; avians; domestic household or farm animals, such as cats,dogs, sheep, goats, cattle, horses and pigs; laboratory animals, such asmice, rats and guinea pigs; fish; reptiles; zoo and wild animals; andthe like.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” thus mean and include, without limitation,antibiotics, anti-arrhythmic agents, anti-viral agents, analgesics,steroidal anti-inflammatories, non-steroidal anti-inflammatories,anti-neoplastics, anti-spasmodics, modulators of cell-extracellularmatrix interactions, proteins, hormones, growth factors, matrixmetalloproteinases (MMPs), enzymes and enzyme inhibitors, anticoagulantsand/or anti-thrombic agents, DNA, RNA, modified DNA and RNA, NSAIDs,inhibitors of DNA, RNA or protein synthesis, polypeptides,oligonucleotides, polynucleotides, nucleoproteins, compounds modulatingcell migration, compounds modulating proliferation and growth of tissue,and vasodilating agents.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” thus include, without limitation, atropine,tropicamide, dexamethasone, dexamethasone phosphate, betamethasone,betamethasone phosphate, prednisolone, triamcinolone, triamcinoloneacetonide, fluocinolone acetonide, anecortave acetate, budesonide,cyclosporine, FK-506, rapamycin, ruboxistaurin, midostaurin,flurbiprofen, suprofen, ketoprofen, diclofenac, ketorolac, nepafenac,lidocaine, neomycin, polymyxin b, bacitracin, gramicidin, gentamicin,oyxtetracycline, ciprofloxacin, ofloxacin, tobramycin, amikacin,vancomycin, cefazolin, ticarcillin, chloramphenicol, miconazole,itraconazole, trifluridine, vidarabine, ganciclovir, acyclovir,cidofovir, ara-amp, foscarnet, idoxuridine, adefovir dipivoxil,methotrexate, carboplatin, phenylephrine, epinephrine, dipivefrin,timolol, 6-hydroxydopamine, betaxolol, pilocarpine, carbachol,physostigmine, demecarium, dorzolamide, brinzolamide, latanoprost,sodium hyaluronate, insulin, verteporfin, pegaptanib, ranibizumab, andother antibodies, antineoplastics, anti-VEGFs, ciliary neurotrophicfactor, brain-derived neurotrophic factor, bFGF, Caspase-1 inhibitors,Caspase-3 inhibitors, α-Adrenoceptors agonists, NMDA antagonists, Glialcell line-derived neurotrophic factors (GDNF), pigmentepithelium-derived factor (PEDF), and NT-3, NT-4, NGF, IGF-2.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” further mean and include the following Class I-ClassV anti-arrhythmic agents: (Class Ia) quinidine, procainamide anddisopyramide; (Class Ib) lidocaine, phenytoin and mexiletine; (Class Ic)flecainide, propafenone and moricizine; (Class II) propranolol, esmolol,timolol, metoprolol and atenolol; (Class III) amiodarone, sotalol,ibutilide and dofetilide; (Class IV) verapamil and diltiazem and (ClassV) adenosine and digoxin.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” further mean and include, without limitation, thefollowing antibiotics: aminoglycosides, cephalosporins, chloramphenicol,clindamycin, erythromycins, fluoroquinolones, macrolides, azolides,metronidazole, penicillins, tetracyclines, trimethoprim-sulfamethoxazoleand vancomycin.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” further include, without limitation, the followingsteroids: andranes (e.g., testosterone), cholestanes, cholic acids,corticosteroids (e.g., dexamethasone), estraenes (e.g., estradiol) andpregnanes (e.g., progesterone).

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” can further include one or more classes of narcoticanalgesics, including, without limitation, morphine, codeine, heroin,hydromorphone, levorphanol, meperidine, methadone, oxycodone,propoxyphene, fentanyl, methadone, naloxone, buprenorphine, butorphanol,nalbuphine and pentazocine.

The terms “pharmacological agent”, “active agent”, “drug” and “activeagent formulation” can further include one or more classes of topical orlocal anesthetics, including, without limitation, esters, such asbenzocaine, chloroprocaine, cocaine, cyclomethycaine,dimethocaine/larocaine, piperocaine, propoxycaine, procaine/novacaine,proparacaine, and tetracaine/amethocaine. Local anesthetics can alsoinclude, without limitation, amides, such as articaine, bupivacaine,cinchocaine/dibucaine, etidocaine, levobupivacaine,lidocaine/lignocaine, mepivacaine, prilocaine, ropivacaine, andtrimecaine. Local anesthetics can further include combinations of theabove from either amides or esters.

The terms “anti-inflammatory” and “anti-inflammatory agent” are alsoused interchangeably herein, and mean and include a “pharmacologicalagent” and/or “active agent formulation”, which, when a therapeuticallyeffective amount is administered to a subject, prevents or treats bodilytissue inflammation i.e. the protective tissue response to injury ordestruction of tissues, which serves to destroy, dilute, or wall offboth the injurious agent and the injured tissues.

Anti-inflammatory agents thus include, without limitation, alclofenac,alclometasone dipropionate, algestone acetonide, alpha amylase,amcinafal, amcinafide, amfenac sodium, amiprilose hydrochloride,anakinra, anirolac, anitrazafen, apazone, balsalazide disodium,bendazac, benoxaprofen, benzydamine hydrochloride, bromelains,broperamole, budesonide, carprofen, cicloprofen, cintazone, cliprofen,clobetasol propionate, clobetasone butyrate, clopirac, cloticasonepropionate, cormethasone acetate, cortodoxone, decanoate, deflazacort,delatestryl, depo-testosterone, desonide, desoximetasone, dexamethasonedipropionate, diclofenac potassium, diclofenac sodium, diflorasonediacetate, diflumidone sodium, diflunisal, difluprednate, diftalone,dimethyl sulfoxide, drocinonide, endrysone, enlimomab, enolicam sodium,epirizole, etodolac, etofenamate, felbinac, fenamole, fenbufen,fenclofenac, fenclorac, fendosal, fenpipalone, fentiazac, flazalone,fluazacort, flufenamic acid, flumizole, flunisolide acetate, flunixin,flunixin meglumine, fluocortin butyl, fluorometholone acetate,fluquazone, flurbiprofen, fluretofen, fluticasone propionate,furaprofen, furobufen, halcinonide, halobetasol propionate, halopredoneacetate, ibufenac, ibuprofen, ibuprofen aluminum, ibuprofen piconol,ilonidap, indomethacin, indomethacin sodium, indoprofen, indoxole,intrazole, isoflupredone acetate, isoxepac, isoxicam, ketoprofen,lofemizole hydrochloride, lomoxicam, loteprednol etabonate,meclofenamate sodium, meclofenamic acid, meclorisone dibutyrate,mefenamic acid, mesalamine, meseclazone, mesterolone,methandrostenolone, methenolone, methenolone acetate, methylprednisolonesuleptanate, momiflumate, nabumetone, nandrolone, naproxen, naproxensodium, naproxol, nimazone, olsalazine sodium, orgotein, orpanoxin,oxandrolane, oxaprozin, oxyphenbutazone, oxymetholone, paranylinehydrochloride, pentosan polysulfate sodium, phenbutazone sodiumglycerate, pirfenidone, piroxicam, piroxicam cinnamate, piroxicamolamine, pirprofen, prednazate, prifelone, prodolic acid, proquazone,proxazole, proxazole citrate, rimexolone, romazarit, salcolex,salnacedin, salsalate, sanguinarium chloride, seclazone, sermetacin,stanozolol, sudoxicam, sulindac, suprofen, talmetacin, talniflumate,talosalate, tebufelone, tenidap, tenidap sodium, tenoxicam, tesicam,tesimide, testosterone, testosterone blends, tetrydamine, tiopinac,tixocortol pivalate, tolmetin, tolmetin sodium, triclonide,triflumidate, zidometacin, and zomepirac sodium.

The term “pharmacological composition”, as used herein, means andincludes a composition comprising a “pharmacological agent” and/or a“biologically active agent” and/or any additional agent or componentidentified herein.

The term “ECM composition”, as used herein, means and includes acomposition comprising at least one ECM.

The term “therapeutically effective”, as used herein, means that theamount of the “pharmacological composition” and/or “pharmacologicalagent” and/or “biologically active agent” administered is of sufficientquantity to ameliorate one or more causes, symptoms, or sequelae of adisease or disorder. Such amelioration only requires a reduction oralteration, not necessarily elimination, of the cause, symptom, orsequelae of a disease or disorder.

The terms “prevent” and “preventing” are used interchangeably herein,and mean and include reducing the frequency or severity of a disease orcondition. The term does not require an absolute preclusion of thedisease or condition. Rather, this term includes decreasing the chancefor disease occurrence.

The terms “treat” and “treatment” are used interchangeably herein, andmean and include medical management of a patient with the intent tocure, ameliorate, stabilize, or prevent a disease, pathologicalcondition, or disorder. The terms include “active treatment”, i.e.treatment directed specifically toward the improvement of a disease,pathological condition, or disorder, and “causal treatment”, i.e.treatment directed toward removal of the cause of the associateddisease, pathological condition, or disorder.

The terms “treat” and “treatment” further include “palliativetreatment”, i.e. treatment designed for the relief of symptoms ratherthan the curing of the disease, pathological condition, or disorder,“preventative treatment”, i.e. treatment directed to minimizing orpartially or completely inhibiting the development of the associateddisease, pathological condition, or disorder, and “supportivetreatment”, i.e. treatment employed to supplement another specifictherapy directed toward the improvement of the associated disease,pathological condition, or disorder.

The terms “optional” and “optionally” mean that the subsequentlydescribed event, circumstance, or material may or may not occur or bepresent, and that the description includes instances where the event,circumstance, or material occurs or is present and instances where itdoes not occur or is not present.

The terms “patient” and “subject” are used interchangeably herein, andmean and include warm blooded mammals, humans and primates; avians;domestic household or farm animals, such as cats, dogs, sheep, goats,cattle, horses and pigs; laboratory animals, such as mice, rats andguinea pigs; fish; reptiles; zoo and wild animals; and the like.

The term “comprise” and variations of the term, such as “comprising” and“comprises,” means “including, but not limited to” and is not intendedto exclude, for example, other additives, components, integers or steps.

The following disclosure is provided to further explain in an enablingfashion the best modes of performing one or more embodiments of thepresent invention. The disclosure is further offered to enhance anunderstanding and appreciation for the inventive principles andadvantages thereof, rather than to limit in any manner the invention.The invention is defined solely by the appended claims including anyamendments made during the pendency of this application and allequivalents of those claims as issued.

In overview, the present disclosure is directed to non-antigenic,resilient, biodegradable biological prostheses, i.e. bioremodelablevascular casted constructs, which are reinforced via perfusion of abiomaterial composition in the vasculature of the construct.

In some embodiments of the invention, the ECM member comprises aseamless tubular ECM member comprising ECM material derived from amammalian tissue source.

According to the invention, the seamless tubular members of theinvention can comprise any mammalian tubular structure, including,without limitation, a segment of a large or small intestine, umbilicalartery or vein, ureter, mesenteric vessel and jugular vein.

According to the invention, the seamless member can thus comprise,without limitation, SIS, SS, gastrointestinal extracellular matrix andumbilical cord extracellular matrix.

In some embodiments, the seamless member can also comprise mesothelialtissue.

In some embodiments, the ECM member comprises a planar ECM membercomprising ECM material derived from a mammalian tissue source.

In some embodiments, the planar ECM member comprises a seamed tubularECM member comprising ECM material derived from a mammalian tissuesource.

According to the invention, the planar and tubular member mammaliantissue sources include, without limitation, the small intestine, largeintestine, stomach, lung, liver, kidney, pancreas, placenta, heart,bladder, prostate, tissue surrounding growing enamel, tissue surroundinggrowing bone, and any fetal tissue from any mammalian organ.

The planar and tubular member (seamless and seamed) ECM material canthus comprise ECM selected from the group comprising, withoutlimitation, SIS, UBS, SS, central nervous system tissue, dermalextracellular matrix, subcutaneous extracellular matrix,gastrointestinal extracellular matrix, tissue surrounding growing bone,placental extracellular matrix, omentum extracellular matrix,mesothelial tissue, cardiac extracellular matrix, kidney extracellularmatrix, pancreas extracellular matrix, lung extracellular matrix, andcombinations thereof.

In some embodiments, the planar and tubular member ECM comprisesmesothelial tissue.

In a preferred embodiment, the casted construct, i.e. planar or tubularmember, mammalian tissue source comprises an adolescent mammalian tissuesource, i.e. an adolescent mammal, such as a piglet, which is preferablyless than three (3) years of age.

The casted construct ECM material can also be derived from the same ordifferent mammalian tissue sources, as disclosed in Co-Pendingapplication Ser. Nos. 13/033,053 and 13/033,102; which are incorporatedby reference herein.

According to the invention, the casted construct ECM material can beused in whole or in part, so that, for example, an ECM material cancontain just the basement membrane (or transitional epithelial layer)with the subadjacent tunica propria, the tunica submucosa, tunicamuscularis, and tunica serosa. The ECM material component of thecomposition can contain any or all of these layers, and thus couldconceivably contain only the basement membrane portion, excluding thesubmucosa. However, generally, and especially since the submucosa isthought to contain and support the active growth factors and otherproteins necessary for in vivo tissue regeneration, the ECM or matrixcomposition from any given source will contain the active extracellularmatrix portions that support cell development and differentiation andtissue regeneration.

In some embodiments, the casted construct ECM material comprises thesubmucosal layer.

In some embodiments, the casted construct ECM material comprises theepithelial basement membrane.

In some embodiments, the casted construct ECM material comprises thesubmucosal and the mucosal layers further comprising the muscularismucosa therebetween.

In some embodiments, the casted construct ECM material comprises thesubmucosal, mucosal and muscularis layers.

In some embodiments the casted construct ECM material comprises thesubmucosal, mucosal, muscularis and serosa layers.

According to the invention, any of the aforementioned layers of thecasted construct ECM material can be delaminated to accommodate variousstructures and applications.

In a preferred embodiment, the casted construct ECM Material comprisessterilized acellular ECM material.

According to the invention, the casted construct ECM material can besterilized via applicant's proprietary Novasterilis® processes disclosedin U.S. Pat. No. 7,108,832 and U.S. patent application Ser. Nos.13/267,337 and 13/480,205; which are incorporated by reference herein intheir entirety.

As set forth in U.S. application Ser. No. 13/480,205, additionalbiologically active and pharmacological agents can be disposed on and/orincorporated (or diffused) into the casted ECM constructs of theinvention.

According to the invention, the casted construct ECM material can alsobe sterilized via the perfusion of a sterilant delivered into thevasculature.

In some embodiments, the sterilant comprises a sterilant compositioncomprising at least one sterilant.

According to the invention, suitable sterilants include, withoutlimitation, acetic acid and chemical derivatives thereof, peraceticacid, trifluoroacetic acid, hydrogen peroxide, glutaraldehyde andcombinations thereof.

In some embodiments, the sterilant composition comprises a Sporeclenz®sterilant, i.e. a mixture comprising acetic acid, hydrogen peroxide, andperacetic acid.

Preferably, the Sporeclenz® sterilant composition is delivered into thevasculature in a sterilization-enhancing effective amount in the rangeof approximately 0.001-2.0 vol. %.

In some embodiments, the sterilant composition comprises a peraceticacid composition further comprising an organic alcohol including,without limitation, ethyl alcohol, isopropyl alcohol and methanol.

Preferably, the alcohol concentration of the peracetic acid compositionis in the range of approximately 1-99.9 vol. %, more preferably, in therange of approximately 5-20 vol. %.

In some embodiments, the sterilant composition comprises at least onedetergent.

According to the invention, suitable detergents include, withoutlimitation, non-ionic detergents selected from the group comprisingzwitter ionic detergents, deoxycholic acid, triton X-100, Tween 20, brij58, brij 96, brij 98, lubrol WX, CHAPS(3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) andcombinations thereof.

In a preferred embodiment, the casted construct ECM material vasculatureis perfused with a detergent composition prior to perfusion with atleast one additional sterilant composition, such as peracetic acid.

Applicant has found that the perfusion of a detergent throughout thevasculature prior to perfusion provides at least 98% removal of raw DNAcontent compared to 94% removal of raw DNA content from treatmentcomprising the perfusion of a sterilant/alcohol composition prior totreatment with a detergent composition.

In some embodiments, the casted construct ECM material vasculature isperfused with at least one additional sterilant composition, such as, byway of example, peracetic acid, prior to perfusion with the detergentcomposition.

In some embodiments, the casted construct ECM material is incubated for24 to 48 hours in 0.5-1% Triton X-100 and 0.5-1% Deoxycholic acid with 5mM EDTA in Dulbecco's Phosphate Buffered Saline (DPBS). In this aspect,it is contemplated that flat or sheet-like casted construct ECMmaterials, such as stomach submucosa (SS), small intestine submucosa(SIS), and bladder submucosa (UBS), can be incubated in a stretchedconfiguration. It is further contemplated that the casted construct ECMmember (or material) vasculature can be perfused with the variousdisclosed sterilant compositions by use of a peristaltic pump.

In some embodiments, the sterilant composition is removed via washingwith a buffer composition comprising at least one buffer.

According to the invention, suitable buffers include, withoutlimitation, phosphate buffered saline (PBS), Earle's balanced saltsolution (EBSS), Hank's balanced salt solution (HBSS), and Dulbecco'sphosphate buffered saline (DPBS).

In some embodiments, the casted construct ECM material is rinsed with abuffer up to six times, including one, two, three, four, five, or sixtimes, with each rinse lasting for about thirty minutes. In an exemplaryaspect, it is contemplated that the step of rinsing the casted constructECM material can comprise rinsing the casted construct ECM materialthree times, with each rinse lasting for about thirty minutes.

In some embodiments, the buffer composition wash is perfused throughoutthe casted construct ECM member (or material) vasculature via aperistaltic pump.

In some embodiments, the casted construct ECM vasculature is perfusedwith a crosslinking composition comprising at least one crosslinkingagent.

According to the invention, suitable crosslinking agents include,without limitation, glutaraldehyde, formaldehyde, polyepoxides,diisocyanates, acyl azides, and the aforementioned alcohols andcombinations thereof.

In some embodiments, the casted construct ECM material is crosslinked inthe range of approximately 0.1-100%.

In some embodiments, the crosslinking agent is continuously perfused inthe vasculature for a period of time in the range of approximately 1-48hours.

In some embodiments, the biomaterial composition disposed within thevasculature of an ECM member comprises at least one ECM-mimickingbiomaterial composition.

In some embodiments, the ECM-mimicking biomaterial composition comprisespoly(glycerol sebacate) (PGS).

Applicant has found that PGS exhibits numerous beneficial propertiesthat provide several beneficial biochemical actions or activities. Theproperties and beneficial actions resulting therefrom are discussed indetail below.

PGS Physical Properties

PGS is a condensate of the non-immunogenic compositions glycerol (asimple sugar alcohol) and sebacic acid (a naturally occurringdicarboxylic acid), wherein, glycerol and sebacic acid are readilymetabolized when proximate mammalian tissue. The non-immunogenicproperties substantially limit the acute inflammatory responsestypically associated with other “biocompatible” polymers, such as ePTFE(polytetrafluoroethylene), that are detrimental to bioremodeling andtissue regeneration.

The mechanical properties of PGS are substantially similar to that ofbiological tissue, indeed, the value of the Young's modulus of PGS isbetween that of a ligament (in KPa range) and tendon (in GPa range). Thestrain to failure of PGS is also similar to that of arteries and veins(i.e. over 260% elongation).

The tensile strength of the PGS is at least 0.28±0.004 MPa. The Young'smodulus and elongation are at least 0.122±0.0003 and at least237.8±0.64%, respectively. For applications requiring strongermechanical properties and a slower biodegradation rate, PGS can beblended with poly(ε-caprolactone) (PCL), i.e. a biodegradable elastomer.

ECM Mimicking Properties/Actions

It has been established that PGS induces tissue remodeling andregeneration when administered proximate to damaged tissue, thus,mimicking the seminal regenerative properties of ECM and, hence, an ECMcomposition formed therefrom. The mechanism underlying this behavior isdeemed to be based on the mechanical and biodegradation kinetics of thePGS. See Sant, et al., Effect of Biodegradation and de novo MatrixSynthesis on the Mechanical Properties of VIC-seeded PGS-PCL scaffolds,Acta. Biomater., vol. 9(4), pp. 5963-73 (2013).

In some embodiments, the ECM-mimicking biomaterial composition furthercomprises at least one of the aforementioned ECM materials.

In some embodiments of the invention, the ECM-mimicking biomaterialcomposition comprises PGS and poly(ε-caprolactone) (PCL). According tothe invention, the addition of PCL to the ECM-mimicking biomaterialcomposition enhances the structural integrity and modulates thedegradation of the composition.

In some embodiments, the ECM/ECM-mimicking biomaterial compositionfurther comprises PCL.

In some embodiments, the ECM-mimicking biomaterial composition comprisespoly(glycerol sebacate) acrylate (PGSA), which, according to theinvention, can be crosslinked and/or cured via the combination of aphotoinitiator and radiation.

According to the invention, suitable photoinitiators for radiationinduced crosslinking comprise, without limitation,2-hydroxy-1-[4-hydroxyethoxy)phenyl]-2-methyl-1-propanone (D 2959, CibaGeigy), 2,2-dimethoxy-2-phenylacetophenone, titanocenes, fluorinateddiaryltitanocenes, iron arene complexes, manganese decacarbonyl,methylcyclopentadienyl manganese tricarbonyl and any organometallaticphotoinitiator that produces free radicals or cations.

According to the invention, suitable′radiation wavelengths forcrosslinking and/or curing the ECM-mimicking biomaterial compositioncomprise, without limitation, visible light; particularly, radiation inthe range of approximately 380-750 nm, and ultraviolet (UV) light,particularly, radiation in the range of 10-400 nm, which includesextreme UV (10-121 nm), vacuum UV (10-200 nm), hydrogen lyman α-UV(121-122 nm), Far UV (122-200 nm), Middle UV (200-300 nm), Near UV(300-400 nm), UV-C (100-280 nm), UV-B (280-315 nm) and UV-A (315-400 nm)species of UV light.

In some embodiments, the ECM-mimicking biomaterial composition comprisesa co-polymer of PGSA and polyethylene glycol (PEG) diacrylate.

Preferably, the ratio of PGSA to PEG diacrylate used when developing thephotocured PGSA hydrogel is proportional to the physical strength of thebiomaterial composition, wherein, when the PGSA/PEG diacrylatecomposition has a ratio of PGSA to PEG diacrylate in the range of95:05-50:50, the composition exhibits a Young's modulus in the range ofapproximately 0.5-20 MPa.

According to the invention, the Young's modulus will also vary based onthe configuration of the casted constructs, such as the pattern of thevasculature.

In some embodiments of the invention, the ECM-mimicking biomaterialcomposition comprises an ECM/ECM-mimicking biomaterial composition, e.g.50% ECM/50% PGS.

In some embodiments, the biomaterial composition disposed within thevasculature of an ECM member comprises a polymeric compositioncomprising at least one biocompatible polymeric material.

According to the invention, the biocompatible polymeric material cancomprise, without limitation, polyglycolide (PGA), polylactide (PLA),poly(ε-caprolactone) (PCL), poly dioxanone (a polyether-ester), polylactide-co-glycolide, polyamide esters, polyalkalene esters, polyvinylesters, polyvinyl alcohol, and polyanhydrides, and like polymers.

The biocompatible polymeric material can also comprise, withoutlimitation, natural polymeric materials, including, without limitation,polysaccharides (e.g. starch and cellulose), proteins (e.g., gelatin,casein, silk, wool, etc.), and polyesters (e.g., polyhydroxyalkanoates).

In some embodiments of the invention, the biocompatible polymericmaterial comprises, without limitation, polyhydroxyalkonates (PHAs),polylactides (PLLA) and polyglycolides (PLGA) and their copolymers, forexample poly(ε-caprolactone-co-glycolide), polyanhydrides, and likepolymers.

According to the invention, biocompatible polymeric materials can alsocomprise a hydrogel composition, including, without limitation,polyurethane, poly(ethylene glycol), poly(propylene glycol),poly(vinylpyrrolidone), xanthan, methyl cellulose, carboxymethylcellulose, alginate, hyaluronan, poly(acrylic acid), polyvinyl alcohol,acrylic acid, hydroxypropyl methyl cellulose, methacrylic acid,αβ-glycerophosphate, κ-carrageenan, 2-acrylamido-2-methylpropanesulfonicacid, and β-hairpin peptide.

In some embodiments, the hydrogel composition is crosslinked by anenzymatic composition.

According to the invention, suitable crosslinking enzymatic compositionscomprise, without limitation, transglutaminase, lysyl oxidase andriboflavin.

In some embodiments, the hydrogel composition is crosslinked and/orcured via exposure to radiation.

In some embodiments, the radiation comprises one of the aforementionedradiation wavelengths.

In some embodiments, the hydrogel composition comprises at least oneaforementioned photoinitiator.

In some embodiments, the polymeric composition is plasma treated toaccommodate hygroscopic agents.

In some embodiments, the polymeric composition comprises athermosensitive chitosan hydrogel composition.

In some embodiments, the thermosensitive chitosan composition comprisesa chitosan/glycerophosphate composition.

In some embodiments, the thermosensitive chitosan composition setsand/or gelates at a temperature in the range of 30-40° C.

Chitosan (including a thermosensitive chitosan composition) exhibits awide range of favorable biochemical properties that make it anoutstanding agent for use in the medical field. The biochemicalproperties of chitosan, which are discussed below, includebiocompatibility, biodegradability and non-toxicity. Additionalproperties, such as analgesic, hemostatic, antimicrobial, andantioxidant have also been reported. See Aranaz, et al., FunctionalCharacterization of Chitin and Chitosan, Current Chemical Biology, vol.3, pp. 203-230 (2009); and Kumar MNVR, A Review of Chitin and ChitosanApplications, React. Funct. Polm., vol. 46, pp. 1-27 (2000).

Biodegradability of Chitosan

Although chitosan is absent from mammals, chitosan can be readilydegraded in vivo by several proteases (lysozyme, papain, pepsin, etc.).The biodegradation of chitosan leads to the release of non-toxicoligosaccharides of variable length, which can be subsequentlyincorporated to glycosaminoglycans and glycoproteins, to metabolicpathways or be excreted. See Pangburn, et al., Lysozyme Degradation ofPartially Deacetylated Chitin, its Films and Hydrogels, Biomaterials,vol. 3(2), pp. 105-108 (1982).

Biocompatibility of Chitosan

Chitosan also exhibits very good compatibility; particularly,cytocompatibility. The enhanced cytocompatibility of chitosan has beenproven in vitro with myocardial, endothelial and epithelial cells,fibroblast, hepatocytes, chondrocytes and keratinocytes. See Chatelet,et al., Influence of the Degree of Acetylation on Some BiologicalProperties of Chitosan Films, Biomaterials, vol. 22(3), pp. 261-268(2001).

Analgesic Properties of Chitosan

Chitosan exhibits very favorable analgesic properties (or effects).Okamoto, et al. specifically studied the analgesic effect of chitosan oninflammatory pain due to intraperitoneal administration of acetic acid.See Okamoto, et al., Analgesic Effects of Chitin and Chitosan, Carbohyd.Poly., vol. 49, pp. 249-252 (2002).

Okamoto, et al. found that, due to its polycationic nature, the freeprimary amino groups of chitosan can protonate in the presence of protonions and, thereby, reduce the pH, which is a primary cause of theanalgesic properties. From experimental data, it was also concluded thatthe analgesic effect was due primarily to the absorption of bradykinin,one of the main components (or substances) related to pain.

Hemostatic Properties of Chitosan

Chitosan, as well as sulphated chitosan oligomers, further exhibitsanticoagulant activity. The anticoagulant activity of chitosan is deemedto be related to its positive charge, since red blood cells' membranesare negatively charged. See Rao, et al., Use of Chitosan as Biomaterial:Studies on its Safety and Hemostatic Potential, J. Biomed. Mat. Res.,vol. 34, pp. 21-28 (1997).

Permeation Enhancing Properties of Chitosan

Chitosan also acts as a permeation enhancer by opening epithelial tightjunctions. The mechanism underlying this behavior is deemed to be basedon the interaction of positively charged chitosan and the cell membraneresulting in a reorganization of the tight junction-associated proteins.See Smith, et al., Effect of Chitosan on Epithelial Cell TightJunctions, Pharm. Res., vol. 21(1), pp. 43-49 (2004).

Antimicrobial Properties of Chitosan

Chitosan also exhibits antimicrobial activity against different groupsof microorganisms, such as bacteria, yeast, and fungi. Two mainmechanisms have been suggested as the cause of the inhibition ofmicrobial cells by chitosan.

The first mechanism comprises the interaction with anionic groups on thecell surface due to chitosan's polycationic nature, which causes theformation of an impermeable layer around the cell.

The second mechanism involves the inhibition of the RNA and proteinsynthesis by permeation into the cell nucleus. See Liu et al.,Antibacterial Action of Chitosan and Carboxymethylated Chitosan, J.Appl. Polym. Sci., vol. 79(7), pp. 1324-1335 (2001).

Antioxidative Properties of Chitosan

Chitosan has also shown a significant scavenging capacity againstdifferent radical species; the results being comparable to thoseobtained with commercial antioxidants. See Park, et al., Free RadicalScavenging Activities of Differently Deacetylated Chitosans, Carbohyd.Polym., vol. 55(1), pp. 17-22 (2004).

Tissue Repair of Chitosan

By virtue of the above discussed properties of chitosan, chitosan canand, in most instances, will enhance the repair of damaged tissue.Indeed, it has been found that chitosan activates immunocytes andinflammatory cells, such as PMN, macrophage, fibroblasts and endothelialcells. See Ueno, et al., Topical Formulations and Wound HealingApplications of Chitosan, Adv. Drug Del. Res., vol. 52, pp. 105-115(2001).

Chitosan oligomers have also exhibited tissue repair properties. It hasbeen suggested that the tissue repair properties are due to theirability to stimulate fibroblast production by affecting the basicfibroblast growth factor. Subsequent collagen production furtherfacilitates the formation of connective tissue.

In some embodiments, the biomaterial composition disposed within thevasculature of the ECM member comprises an ECM composition comprising atleast one of the aforementioned ECM materials.

In some embodiments, the ECM material is crosslinked via a crosslinkingagent.

According to the invention, suitable crosslinking agents include,without limitation, glutaraldehyde, formaldehyde, polyepoxides,diisocyanates, acyl azides, and the aforementioned alcohols.

In some embodiments, the ECM material is enzymatically crosslinked.

According to the invention, suitable agents for enzymatic crosslinkingcomprise, without limitation, transglutaminase, lysyl oxidase andriboflavin.

In some embodiments, the ECM material is crosslinked and/or cured viaexposure to radiation.

In some embodiments, the radiation comprises one of the aforementionedwavelengths.

In some embodiments, the casted construct comprises at least onecoating.

Suitable coatings are disclosed in Co-Pending application Ser. Nos.14/566,155, 14/566,306 and 14/566,209, which are incorporated byreference herein in their entirety.

In some embodiments, the coating comprises an ECM composition comprisingat least one of the aforementioned ECM materials.

In some embodiments, the coating comprises a biodegradable polymericcomposition comprising one of the aforementioned polymeric compositions.

In some embodiments, the coating comprises one of the aforementionedECM-mimicking biomaterial compositions.

In some embodiments, the coating comprises one of the aforementionedECM/ECM-mimicking biomaterial compositions.

In some embodiments, the coating comprises a blend of the aforementionedECM and/or polymeric compositions and/or ECM-mimicking biomaterialcompositions and/or ECM/ECM-mimicking biomaterial compositions.

In some embodiments of the invention, the casted constructs furthercomprise an outer reinforcing structure, such as disclosed in Co-pendingU.S. application Ser. No. 14/337,863, filed on Jul. 22, 2014, and Ser.Nos. 14/554,730, 14/554,795 and 14/554,847, filed on Nov. 26, 2014,which are incorporated by reference herein in their entirety.

According to the invention, the reinforcing structure can comprise awound member or strand configuration, i.e. a thin strand wound aroundthe outer surface of the tubular member, such as disclosed in Co-Pendingapplication Ser. No. 14/337,863 or a mesh structure, such as disclosedin Co-Pending application Ser. Nos. 14/554,730, 14/554,795 and14/554,847.

In some embodiments of the invention, the reinforcing structurecomprises a mesh or woven structure.

In some embodiments of the invention, the reinforcing structurecomprises one of the aforementioned ECM materials.

In some embodiments, the reinforcing structure comprises one of theaforementioned polymeric compositions.

In some embodiments of the invention, the reinforcing structurecomprises one of the aforementioned ECM-mimicking biomaterialcompositions.

In some embodiments of the invention, the reinforcing structurecomprises one of the aforementioned ECM/ECM-mimicking biomaterialcompositions.

In some embodiments of the invention, the reinforcing structurecomprises a biocompatible metal, such as stainless steel and Nitinol®.

As indicated above, in some embodiments of the invention, the ECMmember(s) and/or biomaterial composition(s) and/or coating(s) and/orreinforcing structure(s) and, hence, casted construct formed therefromor therewith includes at least one additional biologically active agentor composition, i.e. an agent that induces or modulates a physiologicalor biological process, or cellular activity, e.g., inducesproliferation, and/or growth and/or regeneration of tissue.

Suitable biologically active agents include any of the aforementionedbiologically active agents, including, without limitation, theaforementioned cells, proteins and growth factors.

In some embodiments, the ECM member(s) and/or biomaterial composition(s)and/or coating(s) and/or reinforcing structure(s) and, hence, castedconstruct formed therefrom or therewith includes at least onepharmacological agent or composition (or drug), i.e. an agent orcomposition that is capable of producing a desired biological effect invivo, e.g., stimulation or suppression of apoptosis, stimulation orsuppression of an immune response, etc.

Suitable pharmacological agents and compositions include any of theaforementioned agents, including, without limitation, antibiotics,anti-viral agents, analgesics, steroidal anti-inflammatories,non-steroidal anti-inflammatories, anti-neoplastics, anti-spasmodics,modulators of cell-extracellular matrix interactions, proteins,hormones, enzymes and enzyme inhibitors, anticoagulants and/oranti-thrombic agents, DNA, RNA, modified DNA and RNA, NSAIDs, inhibitorsof DNA, RNA or protein synthesis, polypeptides, oligonucleotides,polynucleotides, nucleoproteins, compounds modulating cell migration,compounds modulating proliferation and growth of cells and/or tissue,and vasodilating agents.

In some embodiments of the invention, the pharmacological agentcomprises a statin, i.e. a HMG-CoA reductase inhibitor. According to theinvention, suitable statins include, without limitation, atorvastatin(Lipitor®), cerivastatin, fluvastatin (Lescol®), lovastatin (Mevacor®,Altocor®, Altoprev®), mevastatin, pitavastatin (Livalo®, Pitava®),pravastatin (Pravachol®, Selektine®, Lipostat®), rosuvastatin(Crestor®), and simvastatin (Zocor®, Lipex®). Several actives comprisinga combination of a statin and another agent, such asezetimbe/simvastatin (Vytorin®), are also suitable.

Applicant has found that the noted statins exhibit numerous beneficialproperties that provide several beneficial biochemical actions oractivities. Among the beneficial biochemical actions, Applicant hasfound that when a statin is added to ECM (wherein a statin augmented ECMmember or casted construct is formed) and the statin augmented ECMmember is administered to damaged tissue, the statin interacts with thecells recruited by the ECM, wherein the statin augmented ECM membermodulates inflammation of the damaged tissue by modulating severalsignificant inflammatory processes, including restricting expression ofmonocyte chemoattractant protein-1 (MCP-1) and chemokine (C-C) motifligand 2 (CCR2).

Further beneficial actions are discussed in detail in Applicant'sCo-Pending application Ser. No. 13/328,287, filed on Dec. 16, 2011, Ser.No. 13/373,569, filed on Sep. 24, 2012 and Ser. No. 13/782,024, filed onMar. 1, 2013; which are incorporated by reference herein in theirentirety.

Additional suitable pharmacological agents and compositions that can bedelivered within the scope of the invention are disclosed in Pat. Pub.Nos. 20070014874, 20070014873, 20070014872, 20070014871, 20070014870,20070014869, and 20070014868; which are expressly incorporated byreference herein in its entirety.

In some embodiments of the invention, the biologically active agentcomprises a protein selected from the group comprising, withoutlimitation, collagen (types I-V), proteoglycans, glycosaminoglycans(GAGS), glycoproteins, cytokines, cell-surface associated proteins, andcell adhesion molecules (CAMs).

In some embodiments, the biologically active agent provides a structuralsupport scaffold. Suitable bioactive agents include, without limitation,elastin and ECM having additional GAG content, such as additionalhyaluronic acid and/or chondroitin sulfate.

According to the invention, the biologically active and pharmacologicalagents referenced above can comprise various forms. In some embodimentsof the invention, the biologically active and pharmacological agents,e.g. simvastatin, comprise microcapsules that provide delayed deliveryof the agent contained therein.

In some embodiments, the ECM member(s) and/or biomaterial composition(s)and/or coating(s) and/or reinforcing structure(s) and, hence, castedconstruct formed therefrom or therewith provides a single-stage agentdelivery profile, i.e. comprise a single-stage delivery vehicle, whereina modulated dosage of an aforementioned biologically active and/orpharmacological agent is provided.

According to the invention, the term “modulated dosage” as used herein,and variants of this language generally refer to the modulation (e.g.,alteration, delay, retardation, reduction, etc.) of a process involvingdifferent eluting or dispersal rates of an agent within biologicaltissue.

In some embodiments, the single-stage delivery vehicle comprisesencapsulated particulates of a biologically active and/orpharmacological agent.

In some embodiments, the encapsulation composition comprises one of theaforementioned ECM compositions.

In some embodiments, the encapsulation composition comprises one of theaforementioned polymeric compositions.

In some embodiments, the encapsulation composition comprises one of theaforementioned ECM-mimicking biomaterial compositions.

In some embodiments, the encapsulation composition comprises one of theaforementioned ECM/ECM-mimicking biomaterial compositions.

In some embodiments, the encapsulation composition comprises an osmoticfluctuation inducing composition. According to the invention, suitableosmotic fluctuation inducing compositions include, without limitation,PEG, alginate and dextran.

According to the invention, the term “osmotic fluctuation” as usedherein, and variants of this language generally refer to the modulationof the osmotic pressure gradient across a defined barrier.

For example, as is well known in the art, alginate is capable ofabsorbing 200-300 times its weight in water, which substantiallyincreases the osmotic pressure gradient of the alginate. The increasedosmotic pressure gradient of the alginate results in a rapid dispersalof an agent therefrom.

In some embodiments of the invention, the ECM member(s) and/orbiomaterial composition(s) and/or coating(s) and/or reinforcingstructure(s) and, hence, casted construct formed therefrom or therewithprovides a multi-stage agent delivery profile, i.e. comprise amulti-stage agent delivery vehicle, wherein a plurality of theaforementioned biologically active and/or pharmacological agents areadministered via a modulated dosage. By way of example, in someembodiments, the multi-stage delivery vehicle comprises encapsulatedparticulates comprising an antibiotic composition encapsulated in analginate composition having a statin incorporated therein, whichprovides a tiered modulated agent delivery.

In some embodiments, the multi-stage agent delivery vehicle comprises acombination of different biologically active and/or pharmacologicalagents. By way of example, in some embodiments, the multi-stage deliveryvehicle comprises encapsulated particulates comprising an encapsulatedgrowth factor concomitantly administered with an encapsulatedanti-inflammatory.

In some embodiments, the multi-stage delivery vehicle comprises aplurality of different biologically active and/or pharmacological agentsencapsulated in different encapsulation compositions. By way of example,in some embodiments, the multi-stage delivery vehicle comprisesencapsulated particulates comprising a growth factor encapsulated inalginate composition and a pharmacological agent encapsulated in apolyglycolide composition.

According to the invention and indicated above, upon disposing a castedconstruct of the invention proximate damaged or diseased biologicaltissue, “modulated healing” is effectuated.

The term “modulated healing”, as used herein, and variants of thislanguage generally refer to the modulation (e.g., alteration, delay,retardation, reduction, etc.) of a process involving different cascadesor sequences of naturally occurring tissue repair in response tolocalized tissue damage or injury, substantially reducing theirinflammatory effect. Modulated healing, as used herein, includes manydifferent biologic processes, including epithelial growth, fibrindeposition, platelet activation and attachment, inhibition,proliferation and/or differentiation, connective fibrous tissueproduction and function, angiogenesis, and several stages of acuteand/or chronic inflammation, and their interplay with each other.

For example, in some embodiments, the ECM member and/or biomaterialcomposition and/or coating and/or reinforcing structure and, hence,casted construct formed therefrom or therewith is specificallyformulated (or designed) to alter, delay, retard, reduce, and/or detainone or more of the phases associated with healing of damaged tissue,including, but not limited to, the inflammatory phase (e.g., platelet orfibrin deposition), and the proliferative phase when in contact withbiological tissue.

In some embodiments of the invention, “modulated healing” means andincludes the ability of ECM member(s) and/or biomaterial composition(s)and/or coating(s) and/or reinforcing structure(s) and, hence, castedconstructs formed therewith to restrict the expression of inflammatorycomponents. By way of example, according to the invention, when an ECMmember and/or biomaterial composition and/or coating and/or reinforcingstructure and, hence, casted construct formed therewith comprises astatin augmented ECM composition, i.e. a composition comprising an ECMand an exogenously added statin, and the casted construct is disposedproximate damaged biological tissue, the casted construct restrictsexpression of monocyte chemoattractant protein-1 (MCP-1) and chemokine(C-C) motif ligand 2 (CCR2).

In some embodiments, “modulated healing” means and includes the abilityof an ECM member and/or biomaterial composition and/or coating and/orreinforcing structure and, hence, casted construct formed therefrom ortherewith to alter a substantial inflammatory phase (e.g., platelet orfibrin deposition) at the beginning of the tissue healing process. Asused herein, the phrase “alter a substantial inflammatory phase” refersto the ability of a casted construct to substantially reduce theinflammatory response at an injury site when in contact with biologicaltissue.

In such an instance, a minor amount of inflammation may ensue inresponse to tissue injury, but this level of inflammation response,e.g., platelet and/or fibrin deposition, is substantially reduced whencompared to inflammation that takes place in the absence of a castedconstruct of the invention.

The term “modulated healing” also refers to the ability of an ECM memberand/or biomaterial composition and/or coating and/or reinforcingstructure and, hence, casted construct formed therewith to induce hostcell and/or tissue proliferation, bioremodeling, includingneovascularization, e.g., vasculogenesis, angiogenesis, andintussusception, and regeneration of tissue structures withsite-specific structural and functional properties.

Thus, in some embodiments, the term “modulated healing” means andincludes the ability of ECM member(s) and/or biomaterial composition(s)and/or coating(s) and/or reinforcing structure(s) and, hence, castedconstructs formed therewith to modulate inflammation and/or induce hostcell and/or tissue proliferation and remodeling. Again, by way ofexample, according to the invention, when an ECM member and/orbiomaterial composition and/or coating and/or reinforcing structure and,hence, casted construct formed therewith comprises a statin augmentedECM composition, i.e. a composition comprising an ECM and an exogenouslyadded statin, and the casted construct is disposed proximate damagedbiological tissue, the stain interacts with cells recruited by the ECM,wherein the casted construct modulates inflammation by, among otheractions, restricting expression of monocyte chemoattractant protein-1(MCP-1) and chemokine (C-C) motif ligand 2 (CCR2) and induces celland/or tissue proliferation, bioremodeling and regeneration of tissuestructures with site-specific structural and functional properties.

By way of a further example, according to the invention, when a castedconstruct comprises a growth factor augmented ECM composition, i.e. acomposition comprising an ECM and an exogenously added growth factor,e.g. TGF-β, is disposed proximate damaged biological tissue, the growthfactor similarly interacts with the ECM and cells recruited by the ECM,wherein the casted construct modulates inflammation and induces celland/or tissue proliferation, bioremodeling and regeneration of tissue.

In some embodiments, when a casted construct is disposed proximatebiological tissue modulated healing is effectuated through thestructural features of the casted construct. The structural featuresprovide the spatial temporal and mechanical cues to modulate cellpolarity and alignment. The structural features further modulate cellproliferation, migration and differentiation thus modulating the healingprocess.

In some embodiments, the casted constructs provide spatial temporal andmechanical cues.

In some embodiments, the casted constructs are incubated in a maceratingcomposition to degrade the ECM member.

According to the invention, suitable macerating compositions include,without limitation, sodium hydroxide (NaOH), Hydrochloric Acid (HCl),Sulfuric Acid (S₂HO₄), Potassium Hydroxide (KOH), chemical acidsolutions having a pH at least lower than 4 and chemical base solutionshaving a pH at least higher than 10.

In some embodiments, the casted constructs are macerated enzymatically.

In some embodiments, the casted constructs are between 0.1-100%macerated.

In some embodiments, the casted constructs are completely macerated toprovide a biomaterial composition alone, wherein the biomaterialcomposition is substantially devoid of the surrounding ECM member.

In some embodiments of the invention, the tensile strength of the castedconstruct is preferably in the range of approximately 200-5000 KPa.

In some embodiments of the invention, the Young's modulus of the castedconstruct is preferably in the range of approximately 30-1000 KPa.

In some embodiments of the invention, the casted construct comprises aporosity in the range of 10-90%.

According to the invention, the biomaterial compositions of theinvention can be delivered into the ECM member vasculature by variousconventional means.

In some embodiments, the biomaterial composition is injected into atleast one artery.

In some embodiments, the biomaterial composition is injected into atleast one venous vessel.

In some embodiments, the venous vessel comprises a blood vessel.

In some embodiments, the venous vessel comprises a lymphatic vessel.

In some embodiments, the injection into a venous vessel is performed viaa retrograde injection, wherein the composition flow direction isoriented to bypass the native valves present in the venous vessels thatobstruct anterograde injection.

In some embodiments, the total volume of the biomaterial composition isadministered in a single continuous injection.

In some embodiments, the total volume of the biomaterial composition isadministered via a plurality of injections.

In some embodiments, the vasculature of the ECM member is perfused via aperistaltic apparatus.

Referring now to FIG. 1, there is shown an illustration of a segment ofthe vasculature 10 of a small intestine. As illustrated in FIG. 1, thevasculature 10 comprises a primary mesentery artery 12 and a pluralityof capillaries 14.

Referring now to FIG. 2, there is shown the small intestine vasculature10 receiving an anterograde injection of an aforementioned biomaterialcomposition from injection means 100.

Referring now to FIG. 3, there is shown one embodiment of a seamlesstubular casted construct 20 of the invention, which, as indicated above,in some embodiments comprises a section of small intestine. Asillustrated in FIG. 3, the tubular construct 20 includes a lumen 22 anda plurality of capillaries 24.

According to the invention, in some embodiments, casted construct 20 cansimilarly comprise various dimensions to accommodate various biologicalstructures and applications.

EXAMPLES

The following examples are provided to enable those skilled in the artto more clearly understand and practice the present invention. Theyshould not be considered as limiting the scope of the invention, butmerely as being illustrated as representative thereof.

Example 1

Referring now to FIG. 4, there is shown a portion 52 of the resectedsection of porcine small intestine 50 shown in FIG. 6. As illustrated inFIG. 4, the small intestine portion 52 includes the mesentery 54 andcapillaries 56.

After resection, the section of the small intestine 50 was processed,i.e. rinsed, with a gentle detergent (0.5% Triton X-100/0.5% SodiumDeoxycholate in 5 mM EDTA in DPBS) for 24 hours and thereafter rinsedthree times in DPBS (as discussed above), via anterograde injection ofthe respective solutions into the primary mesentery artery 60 (see FIG.6). The rinses were followed by an additional anterograde injection of asterilant composition (2.0% peracetic acid in 5% ethyl alcohol) into themesentery artery 60.

The addition of the sterilant was followed by an additional rinse step(three washes in DPBS).

Referring now to FIG. 5, the additional rinse step was followed by ananterograde injection of a tracing composition (0.4% Trypan Blue) intoprimary mesentery artery 60. As illustrated in FIGS. 5 and 6, thetracing composition was visibly present in the submucosa capillaries 55and the mesentery capillaries 56.

The resultant structure thus comprised a sterilized and acellularsection of porcine small intestine 50 (see FIG. 6).

The example thus confirms that biological tissue can be sterilized anddecellularized via the anterograde injection of detergent and sterilantcompositions into the vasculature.

The example further confirms that a biomaterial composition can and willperfuse into the vasculature of biological tissue.

Example 2

Referring now to FIG. 7, a segment 70 of the sterilized and acellularsection of porcine small intestine described in Example 1 was used toform a tubular seamless casted construct. As indicated above, theporcine small intestine segment 70 can be trimmed to any length toaccommodate various applications.

Example 3

A segment of rabbit small intestine was resected and processed in amanner similar to the porcine small intestine in Example 1. Referringnow to FIG. 8, the vasculature 82 of the sterilized and acellularsection of rabbit small intestine 80 was then infused with a polymericcomposition of the invention (shown in yellow); in this instance, apolymeric composition comprising polyurethane.

The resultant structure 84 comprised a sterilized and acellular ECMmember with a biomaterial composition perfused within the vasculature82.

According to the invention, the casted constructs 70, 84 described aboveand shown in FIGS. 7 and 8, when disposed proximate damaged tissue,modulate inflammation of the damaged tissue by, among other actions,restricting expression of MCP-1 and CCR2, and induce cell and/or tissueproliferation, bioremodeling and regeneration of tissue structures withsite-specific structural and functional properties.

It is to be understood that the casted constructs discussed herein andshown in FIGS. 1-7 are merely examples of the various casted constructsthat can be employed within the scope of the invention. The castedconstructs shown in FIGS. 1-7 should thus not be construed as limitingthe scope of the invention in any manner.

As will be readily appreciated by one having ordinary skill in the art,the casted constructs of the invention can be configured in a variety ofshapes and readily employed in various medical procedures, including,without limitation, treatment of coronary and peripheral vasculardisease (PVD) in cardiovascular vessels, including, but not limited to,iliacs, superficial femoral artery, renal artery, tibial artery,popliteal artery, etc., deep vein thromboses (DVT), vascular bypasses,and coronary vascular repair.

The casted constructs of the invention can also be configured in avariety of shapes and used to repair, augment, reconstruct or replaceother damaged or diseased biological structures and associated tissue,including a pericardium, myocardium, esophagus, trachea, bronchus,ureter, urethra, bile duct, and small and large intestine. The castedconstructs can also be readily employed to reconstruct or replacedamaged or diseased dura around a spinal cord.

The casted constructs can also be readily employed to form a biomaterialpouch configured to encase an ECM or pharmacological composition, ormedical instrument or device, such as a pacemaker, therein. Illustrativepouch configurations are disclosed in U.S. Pat. No. 8,758,448 andApplicant's Co-pending U.S. application Ser. Nos. 13/573,566 and13/896,424, which are incorporated by reference herein in theirentirety.

The casted constructs can also be readily employed to construct a valveconduit that is configured to replace and/or regenerate a damaged and/ordefective heart valve. Illustrative valve configurations are disclosedin U.S. Pat. Nos. 8,696,744 and 8,709,076 and Applicant's Co-pendingU.S. application Ser. No. 13/804,683, which are incorporated byreference herein in their entirety.

The casted constructs can also be readily employed to construct a valveconduit that is configured to replace a damaged and/or defectivevascular structures. Illustrative vascular prostheses are disclosed inU.S. Pat. No. 8,808,363 and Applicant's Co-pending U.S. application Ser.Nos. 14/031,520, 14/337,915 and 14/337,863, which are incorporated byreference herein in their entirety.

One having ordinary skill in the art will thus readily appreciate thatthe casted constructs of the invention provide numerous advantages overconventional apparatus and structures for repairing and/or regeneratingtissue. Among the advantages are the following:

-   -   The provision of casted constructs that can be readily and        effectively employed to treat various damaged or diseased        biological structures and associated tissue;    -   The provision of casted constructs that can be readily employed        to close and maintain closure of openings in biological tissue;    -   The provision of casted constructs that equal or exceed        resilience of prosthetics formed from synthetic compositions,        while being configured to induce host cell and/or tissue        proliferation, bioremodeling and regeneration of new tissue, and        tissue structures with site-specific structural and functional        properties;    -   The provision of casted constructs that provide spatial and        mechanical cues that modulate cell polarity, spatial temporal        positioning, differentiation, proliferation and migration when        in contact with biological tissue; particularly, damaged and/or        diseased tissue cells;    -   The provision of casted constructs that include ECM-mimicking        biomaterials that induce host cell and/or tissue proliferation,        bioremodeling and regeneration of new tissue, and tissue        structures with site-specific structural and functional        properties; and    -   The provision of casted constructs that are configured to        effectively administer at least one biologically active agent        and/or pharmacological agent or composition to a subject's        tissue to induce a desired biological and/or therapeutic effect.

Without departing from the spirit and scope of this invention, one ofordinary skill can make various changes and modifications to theinvention to adapt it to various usages and conditions. As such, thesechanges and modifications are properly, equitably, and intended to be,within the full range of equivalence of any issued claims.

What is claimed is:
 1. A casted construct for treating damagedbiological structures and tissue, comprising: a tissue prosthesiscomprising a reinforced mammalian tissue structure, said mammaliantissue structure comprising first sterilized and acellular extracellularmatrix (ECM) from a first mammalian tissue source, said mammalian tissuestructure further comprising an intact, continuous, reinforced,sterilized and acellular internal vasculature, said internal vasculatureand mammalian tissue structure being sterilized and decellularized byperfusion of a sterilant into said internal vasculature, saidvasculature being infused with an ECM-mimicking biomaterial compositioncomprising poly(glycerol sebacate) (PGS), wherein a PGS augmented tissueprosthesis is formed, said PGS augmented tissue prosthesis being capableof modulating an inflammatory phase of damaged biological tissue, andinducing host cell and tissue proliferation, neovascularization,bioremodeling of said damaged biological tissue and regeneration of newtissue and tissue structures with site-specific structural andfunctional properties, when said PGS augmented tissue prosthesis isdisposed proximate said damaged biological tissue.
 2. The castedconstruct of claim 1, wherein said sterilized first acellular ECMcomprises a crosslinked, sterilized and acellular ECM.
 3. The castedconstruct of claim 1, wherein said first mammalian tissue sourcecomprises a first mammalian tissue selected from the group consisting ofsmall intestine submucosa, urinary bladder submucosa, stomach submucosa,placental tissue, mesothelial tissue, cardiac tissue, kidney tissue,pancreas tissue and lung tissue.
 4. The casted construct of claim 1,wherein said reinforced mammalian tissue structure comprises a planartissue structure.
 5. The casted construct of claim 1, wherein saidreinforced mammalian tissue structure comprises a seamless tubulartissue structure.
 6. The casted construct of claim 5, wherein saidseamless tubular tissue structure comprises a segment of a mammalianstructure selected from the group consisting of mammalian intestine,umbilical artery, umbilical vein, ureter, mesenteric vessel and jugularvein.
 7. The casted construct of claim 5, wherein said seamless tubulartissue structure comprises a segment of adolescent small intestine. 8.The casted construct of claim 1, wherein said ECM-mimicking biomaterialcomposition further comprises poly(ε-caprolactone) (PCL).
 9. The castedconstruct of claim 1, wherein said ECM-mimicking biomaterial compositionfurther comprises second acellular ECM from a second mammalian tissuesource selected from the group consisting of small intestine submucosa,urinary bladder submucosa, stomach submucosa, placental tissue,mesothelial tissue, cardiac tissue, kidney tissue, pancreas tissue andlung tissue.
 10. The casted construct of claim 1, wherein saidECM-mimicking biomaterial composition further comprises a firstbiologically active agent.
 11. The casted construct of claim 10, whereinsaid first biologically active agent comprises a first cell selectedfrom the group consisting of a human embryonic stem cell, fetalcardiomyocyte, myofibroblast and mesenchymal stem cell.
 12. The castedconstruct of claim 10, wherein said first biologically active agentcomprises a first growth factor selected from the group consisting of atransforming growth factor-alpha (TGF-α), transforming growthfactor-beta (TGF-β), basic fibroblast growth factor (bFGF) and vascularepithelial growth factor (VEGF).
 13. The casted construct of claim 1,wherein said ECM-mimicking biomaterial composition further comprises apharmacological agent.
 14. The casted construct of claim 13, whereinsaid pharmacological agent comprises a second statin selected from thegroup consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin,mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin. 15.The casted construct of claim 13, wherein said pharmacological agentcomprises an anti-inflammatory selected from the group consisting ofdesoximetasone, diftalone, flazalone, ibuprofen aluminum, ibuprofen,piconol and rimexolone.
 16. The casted construct of claim 1, whereinsaid reinforced mammalian tissue structure further comprises asupplemental second biologically active agent.
 17. The casted constructof claim 16, wherein said supplemental second biologically active agentcomprises a second cell selected from the group consisting of a humanembryonic stem cell, fetal cardiomyocyte, myofibroblast and mesenchymalstem cell.
 18. The casted construct of claim 16, wherein saidsupplemental second biologically active agent comprises a second growthfactor selected from the group consisting of a transforming growthfactor-alpha (TGF-α), transforming growth factor-beta (TGF-β), basicfibroblast growth factor (bFGF) and vascular epithelial growth factor(VEGF).